E B Aranas, P Z G Fonseca, P F Barker and T S Monteiro
There have been recent rapid developments in stable trapping of levitated nanoparticles in high vacuum. Cooling of nanoparticles, from phonon occupancies of 107 down to $\simeq \,100\mbox{--}1000$ phonons, have already been achieved by several groups. Prospects for quantum ground-state cooling seem extremely promising. Cavity-cooling without added stabilisation by feedback cooling remains challenging, but trapping at high vacuum in a cavity is now possible through the addition of a Paul trap. However, the Paul trap has been found to qualitatively modify the cavity output spectrum, with the latter acquiring an atypical 'split-sideband' structure, of different form from the displacement spectrum, and which depends on N, the optical well at which the particle localises. In the present work we investigate the N-dependence of the dynamics, in particular with respect to thermometry: we show that in strong cooling regions $N\gtrsim 100$, the temperature may still be reliably inferred from the cavity output spectra. We also explain the N-dependence of the mechanical frequencies and optomechanical coupling showing that these may be accurately estimated. We present a simple 'fast-cavity' model for the cavity output and test all our findings against full numerical solutions of the nonlinear stochastic equations of motion for the system.
DOI
Wednesday, April 19, 2017
Tuesday, April 18, 2017
Laser-assisted biofabrication in tissue engineering and regenerative medicine
Sangmo Koo, Samantha M. Santoni, Bruce Z. Gao, Costas P. Grigoropoulos and Zhen Ma
Controlling the spatial arrangement of biomaterials and living cells provides the foundation for fabricating complex biological systems. Such level of spatial resolution (less than 10 µm) is difficult to be obtained through conventional cell processing techniques, which lack the precision, reproducibility, automation, and speed required for the rapid fabrication of engineered tissue constructs. Recently, laser-assisted biofabrication techniques are being intensively developed with the use of computer-aided processes for patterning and assembling both living and nonliving materials with prescribed 2D or 3D organization. In this review, we discuss laser-assisted fabrication methods, including laser tweezers, multi-photon polymerization, laser-induced forward transfer (LIFT), matrix assisted pulsed laser evaporation (MAPLE), and laser ablation as well as their applications in biological science and biomedical engineering. These advanced technologies enable the precise manipulation of in vitro cellular microenvironments and the ability to engineer functional tissue constructs with high complexity and heterogeneity, which serve in regenerative medicine, pharmacology, and basic cell biology studies.
DOI
Controlling the spatial arrangement of biomaterials and living cells provides the foundation for fabricating complex biological systems. Such level of spatial resolution (less than 10 µm) is difficult to be obtained through conventional cell processing techniques, which lack the precision, reproducibility, automation, and speed required for the rapid fabrication of engineered tissue constructs. Recently, laser-assisted biofabrication techniques are being intensively developed with the use of computer-aided processes for patterning and assembling both living and nonliving materials with prescribed 2D or 3D organization. In this review, we discuss laser-assisted fabrication methods, including laser tweezers, multi-photon polymerization, laser-induced forward transfer (LIFT), matrix assisted pulsed laser evaporation (MAPLE), and laser ablation as well as their applications in biological science and biomedical engineering. These advanced technologies enable the precise manipulation of in vitro cellular microenvironments and the ability to engineer functional tissue constructs with high complexity and heterogeneity, which serve in regenerative medicine, pharmacology, and basic cell biology studies.
DOI
Optical Manipulation of Dielectric Nanoparticles with Au Micro-racetrack Resonator by Constructive Interference of Surface Plasmon Waves
Mingrui Yuan, Lin Cheng, Pengfei Cao, Xu Li, Xiaodong He, Xiaoping Zhang
We design a gold micro-racetrack resonator (Au-MRR) which can tightly trap and drive the dielectric nanoparticle to rotate around the circuit of racetrack with an adjustable velocity. Since the surface plasmon waves can be excited and obey the resonance condition of the Au-MRR, the optics force can be strengthened observably due to the resonance. The optical forces applied on dielectric nanoparticle are discussed by utilizing the Maxwell’s stress tensor integration with a numerical finite element method. The depth of longitudinal trapping potential well in the Au-MRR is four times as large as that of a straight waveguide. At the same level of input power, the velocity of particle with radius of 50 nm driven by optical forces on Au-MRR is 200 times larger than that on a straight waveguide. Further, we explore the motion behavior of single nanoparticle lies on different position of Au-MRR, which can provide the details to trap and manipulate multiple nanoparticles and predict their trace of movement. This optimum geometry of Au-MRR allows further enhancement of the optical forces which is expected to realize all-optical on-chip manipulation of nanoparticles, biomolecules, and many other nanomanipulation applications.
DOI
We design a gold micro-racetrack resonator (Au-MRR) which can tightly trap and drive the dielectric nanoparticle to rotate around the circuit of racetrack with an adjustable velocity. Since the surface plasmon waves can be excited and obey the resonance condition of the Au-MRR, the optics force can be strengthened observably due to the resonance. The optical forces applied on dielectric nanoparticle are discussed by utilizing the Maxwell’s stress tensor integration with a numerical finite element method. The depth of longitudinal trapping potential well in the Au-MRR is four times as large as that of a straight waveguide. At the same level of input power, the velocity of particle with radius of 50 nm driven by optical forces on Au-MRR is 200 times larger than that on a straight waveguide. Further, we explore the motion behavior of single nanoparticle lies on different position of Au-MRR, which can provide the details to trap and manipulate multiple nanoparticles and predict their trace of movement. This optimum geometry of Au-MRR allows further enhancement of the optical forces which is expected to realize all-optical on-chip manipulation of nanoparticles, biomolecules, and many other nanomanipulation applications.
DOI
Omnidirectional Transport in Fully Reconfigurable Two Dimensional Optical Ratchets
Alejandro V. Arzola, Mario Villasante-Barahona, Karen Volke-Sepúlveda, Petr Jákl, and Pavel Zemánek
A fully reconfigurable two-dimensional (2D) rocking ratchet system created with holographic optical micromanipulation is presented. We can generate optical potentials with the geometry of any Bravais lattice in 2D and introduce a spatial asymmetry with arbitrary orientation. Nontrivial directed transport of Brownian particles along different directions is demonstrated numerically and experimentally, including on axis, perpendicular, and oblique with respect to an unbiased ac driving. The most important aspect to define the current direction is shown to be the asymmetry and not the driving orientation, and yet we show a system in which the asymmetry orientation of each potential well does not coincide with the transport direction, suggesting an additional symmetry breaking as a result of a coupling with the lattice configuration. Our experimental device, due to its versatility, opens up a new range of possibilities in the study of nonequilibrium dynamics at the microscopic level.
DOI
A fully reconfigurable two-dimensional (2D) rocking ratchet system created with holographic optical micromanipulation is presented. We can generate optical potentials with the geometry of any Bravais lattice in 2D and introduce a spatial asymmetry with arbitrary orientation. Nontrivial directed transport of Brownian particles along different directions is demonstrated numerically and experimentally, including on axis, perpendicular, and oblique with respect to an unbiased ac driving. The most important aspect to define the current direction is shown to be the asymmetry and not the driving orientation, and yet we show a system in which the asymmetry orientation of each potential well does not coincide with the transport direction, suggesting an additional symmetry breaking as a result of a coupling with the lattice configuration. Our experimental device, due to its versatility, opens up a new range of possibilities in the study of nonequilibrium dynamics at the microscopic level.
DOI
Monday, April 17, 2017
Effect of lipopolysaccharide O-side chains on the adhesiveness of Yersinia pseudotuberculosis to J774 macrophages as revealed by optical tweezers
A. A. Byvalov, V. L. KononenkoI. V. Konyshev
A method has been developed for the quantitative estimation of the binding force of a model microsphere with a eukaryocyte based on the optical trap in order to study the molecular mechanism of adhesion between an individual bacterium and a host cell. The substantial role of LPS O-side chains in the adhesiveness of Yersinia pseudotuberculosis 1b to J774 macrophages has been revealed with the use of a set of microspheres functionalized with lipopolysaccharide (LPS) preparations and antibodies with different specificities. The results indicate the significance of the O-antigen as a pathogenicity factor of Y. pseudotuberculosis in colonization of a macroorganism. The developed methodical approaches can be applied to the study of molecular mechanisms of the pathogenesis of pseudotuberculosis and other infectious diseases to improve antiepidemic service.
DOI
A method has been developed for the quantitative estimation of the binding force of a model microsphere with a eukaryocyte based on the optical trap in order to study the molecular mechanism of adhesion between an individual bacterium and a host cell. The substantial role of LPS O-side chains in the adhesiveness of Yersinia pseudotuberculosis 1b to J774 macrophages has been revealed with the use of a set of microspheres functionalized with lipopolysaccharide (LPS) preparations and antibodies with different specificities. The results indicate the significance of the O-antigen as a pathogenicity factor of Y. pseudotuberculosis in colonization of a macroorganism. The developed methodical approaches can be applied to the study of molecular mechanisms of the pathogenesis of pseudotuberculosis and other infectious diseases to improve antiepidemic service.
DOI
Laser-mediated rupture of chlamydial inclusions triggers pathogen egress and host cell necrosis
Markus C. Kerr, Guillermo A. Gomez, Charles Ferguson, Maria C. Tanzer, James M. Murphy, Alpha S. Yap, Robert G. Parton, Wilhelmina M. Huston & Rohan D Teasdale
Remarkably little is known about how intracellular pathogens exit the host cell in order to infect new hosts. Pathogenic chlamydiae egress by first rupturing their replicative niche (the inclusion) before rapidly lysing the host cell. Here we apply a laser ablation strategy to specifically disrupt the chlamydial inclusion, thereby uncoupling inclusion rupture from the subsequent cell lysis and allowing us to dissect the molecular events involved in each step. Pharmacological inhibition of host cell calpains inhibits inclusion rupture, but not subsequent cell lysis. Further, we demonstrate that inclusion rupture triggers a rapid necrotic cell death pathway independent of BAK, BAX, RIP1 and caspases. Both processes work sequentially to efficiently liberate the pathogen from the host cytoplasm, promoting secondary infection. These results reconcile the pathogen's known capacity to promote host cell survival and induce cell death.
DOI
Remarkably little is known about how intracellular pathogens exit the host cell in order to infect new hosts. Pathogenic chlamydiae egress by first rupturing their replicative niche (the inclusion) before rapidly lysing the host cell. Here we apply a laser ablation strategy to specifically disrupt the chlamydial inclusion, thereby uncoupling inclusion rupture from the subsequent cell lysis and allowing us to dissect the molecular events involved in each step. Pharmacological inhibition of host cell calpains inhibits inclusion rupture, but not subsequent cell lysis. Further, we demonstrate that inclusion rupture triggers a rapid necrotic cell death pathway independent of BAK, BAX, RIP1 and caspases. Both processes work sequentially to efficiently liberate the pathogen from the host cytoplasm, promoting secondary infection. These results reconcile the pathogen's known capacity to promote host cell survival and induce cell death.
DOI
Real-time monitoring and visualization of the multi-dimensional motion of an anisotropic nanoparticle
Gi-Hyun Go, Seungjin Heo, Jong-Hoi Cho, Yang-Seok Yoo, MinKwan Kim, Chung-Hyun Park & Yong-Hoon Cho
As interest in anisotropic particles has increased in various research fields, methods of tracking such particles have become increasingly desirable. Here, we present a new and intuitive method to monitor the Brownian motion of a nanowire, which can construct and visualize multi-dimensional motion of a nanowire confined in an optical trap, using a dual particle tracking system. We measured the isolated angular fluctuations and translational motion of the nanowire in the optical trap, and determined its physical properties, such as stiffness and torque constants, depending on laser power and polarization direction. This has wide implications in nanoscience and nanotechnology with levitated anisotropic nanoparticles.
DOI
As interest in anisotropic particles has increased in various research fields, methods of tracking such particles have become increasingly desirable. Here, we present a new and intuitive method to monitor the Brownian motion of a nanowire, which can construct and visualize multi-dimensional motion of a nanowire confined in an optical trap, using a dual particle tracking system. We measured the isolated angular fluctuations and translational motion of the nanowire in the optical trap, and determined its physical properties, such as stiffness and torque constants, depending on laser power and polarization direction. This has wide implications in nanoscience and nanotechnology with levitated anisotropic nanoparticles.
DOI
Mechanistic Basis for the Binding of RGD- and AGDV-Peptides to the Platelet Integrin αIIbβ3
Olga Kononova, Rustem I. Litvinov, Dmitry S. Blokhin, Vladimir V. Klochkov, John W. Weisel, Joel S. Bennett, and Valeri Barsegov
Binding of soluble fibrinogen to the activated conformation of the integrin αIIbβ3 is required for platelet aggregation and is mediated exclusively by the C-terminal AGDV-containing dodecapeptide (γC-12) sequence of the fibrinogen γ chain. However, peptides containing the Arg-Gly-Asp (RGD) sequences located in two places in the fibrinogen Aα chain inhibit soluble fibrinogen binding to αIIbβ3 and make substantial contributions to αIIbβ3 binding when fibrinogen is immobilized and when it is converted to fibrin. Here, we employed optical trap-based nanomechanical measurements and computational molecular modeling to determine the kinetics, energetics, and structural details of cyclic RGDFK (cRGDFK) and γC-12 binding to αIIbβ3. Docking analysis revealed that NMR-determined solution structures of cRGDFK and γC-12 bind to both the open and closed αIIbβ3 conformers at the interface between the αIIb β-propeller domain and the β3 βI domain. The nanomechanical measurements revealed that cRGDFK binds to αIIbβ3 at least as tightly as γC-12. A subsequent analysis of molecular force profiles and the number of peptide−αIIbβ3 binding contacts revealed that both peptides form stable bimolecular complexes with αIIbβ3 that dissociate in the 60–120 pN range. The Gibbs free energy profiles of the αIIbβ3–peptide complexes revealed that the overall stability of the αIIbβ3-cRGDFK complex was comparable with that of the αIIbβ3−γC-12 complex. Thus, these results provide a mechanistic explanation for previous observations that RGD- and AGDV-containing peptides are both potent inhibitors of the αIIbβ3–fibrinogen interactions and are consistent with the observation that RGD motifs, in addition to AGDV, support interaction of αIIbβ3 with immobilized fibrinogen and fibrin.
DOI
Binding of soluble fibrinogen to the activated conformation of the integrin αIIbβ3 is required for platelet aggregation and is mediated exclusively by the C-terminal AGDV-containing dodecapeptide (γC-12) sequence of the fibrinogen γ chain. However, peptides containing the Arg-Gly-Asp (RGD) sequences located in two places in the fibrinogen Aα chain inhibit soluble fibrinogen binding to αIIbβ3 and make substantial contributions to αIIbβ3 binding when fibrinogen is immobilized and when it is converted to fibrin. Here, we employed optical trap-based nanomechanical measurements and computational molecular modeling to determine the kinetics, energetics, and structural details of cyclic RGDFK (cRGDFK) and γC-12 binding to αIIbβ3. Docking analysis revealed that NMR-determined solution structures of cRGDFK and γC-12 bind to both the open and closed αIIbβ3 conformers at the interface between the αIIb β-propeller domain and the β3 βI domain. The nanomechanical measurements revealed that cRGDFK binds to αIIbβ3 at least as tightly as γC-12. A subsequent analysis of molecular force profiles and the number of peptide−αIIbβ3 binding contacts revealed that both peptides form stable bimolecular complexes with αIIbβ3 that dissociate in the 60–120 pN range. The Gibbs free energy profiles of the αIIbβ3–peptide complexes revealed that the overall stability of the αIIbβ3-cRGDFK complex was comparable with that of the αIIbβ3−γC-12 complex. Thus, these results provide a mechanistic explanation for previous observations that RGD- and AGDV-containing peptides are both potent inhibitors of the αIIbβ3–fibrinogen interactions and are consistent with the observation that RGD motifs, in addition to AGDV, support interaction of αIIbβ3 with immobilized fibrinogen and fibrin.
DOI
Two-laser optical tweezers with a blinking beam
Weronika Lamperska, Jan Masajada, Sławomir Drobczyński, Paweł Gusin
We report on a two-laser holographic optical tweezers setup and present its two major advantages over single-laser one. First, the trap stiffness of a weak trapping beam can be measured with a considerable accuracy. Second, a novel method of examining local viscosity of fluid is proposed. Both measurements are performed based on forcing the oscillations of a microscopic polystyrene bead placed between two optical traps. The two beams are generated by separate laser sources and therefore their trapping power can vary. Moreover, a stronger trap ‘blinks’, modulated by an electronic shutter. The blinking frequency can be precisely adjusted to the experimental conditions, which results in high accuracy of the measurements.
DOI
We report on a two-laser holographic optical tweezers setup and present its two major advantages over single-laser one. First, the trap stiffness of a weak trapping beam can be measured with a considerable accuracy. Second, a novel method of examining local viscosity of fluid is proposed. Both measurements are performed based on forcing the oscillations of a microscopic polystyrene bead placed between two optical traps. The two beams are generated by separate laser sources and therefore their trapping power can vary. Moreover, a stronger trap ‘blinks’, modulated by an electronic shutter. The blinking frequency can be precisely adjusted to the experimental conditions, which results in high accuracy of the measurements.
DOI
Onset of particle trapping and release via acoustic bubbles
Yun Chen, Zecong Fang, Brett Merritt, Dillon Strack, Jie Xu and Sungyon Lee
Trapping and sorting of micro-sized objects is one important application of lab on a chip devices, with the use of acoustic bubbles emerging as an effective, non-contact method. Acoustically actuated bubbles are known to exert a secondary radiation force (FSR) on micro-particles and stabilize them on the bubble surface, when this radiation force exceeds the external hydrodynamic forces that act to keep the particles in motion. While the theoretical expression of FSR has been derived by Nyborg decades ago, no direct experimental validation of this force has been performed, and the relationship between FSR and the bubble's ability to trap particles in a given lab on a chip device remains largely empirical. In order to quantify the connection between the bubble oscillation and the resultant FSR, we experimentally measure the amplitude of bubble oscillations that give rise to FSR and observe the trapping and release of a single microsphere in the presence of the mean flow at the corresponding acoustic parameters using an acoustofluidic device. By combining well-developed theories that connect bubble oscillations to the acoustic actuation, we derive the expression for the critical input voltage that leads to particle release into the flow, in good agreement with the experiments.
DOI
Trapping and sorting of micro-sized objects is one important application of lab on a chip devices, with the use of acoustic bubbles emerging as an effective, non-contact method. Acoustically actuated bubbles are known to exert a secondary radiation force (FSR) on micro-particles and stabilize them on the bubble surface, when this radiation force exceeds the external hydrodynamic forces that act to keep the particles in motion. While the theoretical expression of FSR has been derived by Nyborg decades ago, no direct experimental validation of this force has been performed, and the relationship between FSR and the bubble's ability to trap particles in a given lab on a chip device remains largely empirical. In order to quantify the connection between the bubble oscillation and the resultant FSR, we experimentally measure the amplitude of bubble oscillations that give rise to FSR and observe the trapping and release of a single microsphere in the presence of the mean flow at the corresponding acoustic parameters using an acoustofluidic device. By combining well-developed theories that connect bubble oscillations to the acoustic actuation, we derive the expression for the critical input voltage that leads to particle release into the flow, in good agreement with the experiments.
DOI
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